Survey and new insights in the application of PCR‐based molecular markers for identification of HMW‐GS at the Glu‐B1 locus in durum and bread wheat

Wheat, among all cereal grains, possesses unique characteristics conferred by gluten; in particular, high molecular weight glutenin subunits (HMW‐GS) are of considerable interest as they strictly relate to bread‐making quality and contribute to strengthening and stabilizing dough. Thus, the identification of allelic composition, in particular at the Glu‐B1 locus, is very important to wheat quality improvement. Several PCR‐based molecular markers to tag‐specific HMW glutenin genes encoding Bx and By subunits have been developed in recent years. This study provides a survey of the molecular markers developed for the HMW‐GS at the Glu‐B1 locus. In addition, a selection of molecular markers was tested on 31 durum and bread wheat cultivars containing the By8, By16, By9, Bx17, Bx6, Bx14 and Bx17 Glu‐B1 alleles, and a new assignation was defined for the ZSBy9_aF1/R3 molecular marker that was specific for the By20 allele. We believe the results constitute a practical guide for results that might be achieved by these molecular markers on populations and cultivars with high variability at the Glu‐B1 locus.     

Treatment of severe immune-mediated thrombocytopenia with human IV immunoglobulin in 5 dogs

BACKGROUND: Glucocorticoids with or without other immunotherapy are the initial treatment of choice for dogs with severe immune-mediated thrombocytopenia (IMT). The majority of treated dogs will have improvements in platelet counts within 5 to 7 days of starting therapy, but complications from hemorrhage often occur before a response is seen. Human IV immunoglobulin (hIVIG) blocks Fc receptors on mononuclear phagocytic cells in dogs; it is used in people with idiopathic thrombocytopenic purpura. HYPOTHESIS: The purpose of this study was to describe adverse effects and benefit of hIVIG in addition to conventional immunosuppressive therapy in dogs with severe IMT. ANIMALS: Five client-owned dogs with severe primary IMT. METHODS: Case series. The hospital database was searched for dogs with primary IMT treated with hIVIG. RESULTS: No adverse effects were noted during or after hIVIG infusion in any treated dog. Over a 6-month follow-up, all dogs were clinically normal when using conventional immunosuppressive therapy. Human IVIG was administered 3 days after initiation of immunosuppressive therapy in 4 dogs, and, after 2 days, in 1 dog. In all dogs, the mean platelet counts pre- and 24 hours post-hIVIG infusion (0.28-0.76 g/kg) were 2,500/pL and 50,600/microL (62,750/microL for the 4 responders), respectively. One dog failed to respond as promptly to hIVIG (0.34 g/kg), and the platelet count increased to 66,000/microL after 9 days of immunosuppressive therapy. The mean duration of hospitalization post-hIVIG in all 5 dogs was 1.8 days (12 hours for responders), and the mean total length of hospitalization was 4.6 days (3.5 days for responders). Active hemorrhage resolved and no packed red blood cell transfusions were required after hIVIG infusion for responders. CONCLUSIONS AND CLINICAL IMPORTANCE: Human IVIG was well tolerated and appeared to be associated with rapid platelet count recovery and amelioration of clinical signs in most dogs with IMT.     

Progeny Production of the Copepods Pseudodiaptomus euryhalinus and Tisbe monozota in Monospecific and Mixed Cultures

The objective of this research was to culture the calanoid copepod Pseudodiaptomus euryhalinus, Johnson 1939, and the harpacticoid Tisbe monozota, Bowman, 1962, in monospecific and combined cultures (P. euryhalinus : T. monozota at 1:1, 2:1, and 1:2 starting ratios), and to compare the nauplii, copepodite, and adult production. Mean total production ranged from 740.3 ± 137.5 organisms/L (P. euryhalinus) to 884.3 ± 489.7 organisms/L (T. monozota) for monospecific cultures. The 1:1 ratio mixed cultures gave 780.4 ± 155.8 organisms/L, those with the 2:1 and 1:2 P. euryhalinus and T. monozota starting ratios produced 710.1 ± 195.2 and 799.7 ± 232.5 organisms/L, respectively, and there were no significant difference among treatments. All mixed cultures gave significantly lower copepodite productions than the monocultures of each species. In addition, the tendency to a decreased progeny production of the initial females of T. monozota indicates that the outcome of long‐term mixed cultures would production either a high dominance of P. euryhalinus, or monocultures of this species .     

Effect of fenoldopam on renal function after nephrotomy in normal dogs

OBJECTIVE: To evaluate effects of fenoldopam on renal function in normal dogs subjected to bisection nephrotomy. In addition, effects of bisection nephrotomy on renal function in normal dogs were evaluated. STUDY DESIGN: Controlled, randomized, blinded experiment. SAMPLE POPULATION: Sixteen mixed-breed adult dogs. METHODS: Dogs were paired for sex, body weight, and approximate age and assigned to 1 of 2 groups: fenoldopam (F) or placebo (P). Baseline glomerular filtration rate (GFR) based on quantitative renal scintigraphy using (99m)Tc-DTPA, blood urea nitrogen (BUN), serum creatinine (SCr), urinalysis, and urine culture were performed before surgery. Left nephrotomy was performed via median celiotomy. Group F dogs were administered intravenous (IV) fenoldopam (0.1 microg/kg/min) for 90 minutes, whereas group P dogs were administered an equivalent volume of saline (0.9 % NaCl) solution for 90 minutes. Temperature, heart rate, respiration, direct arterial blood pressure, and urine volume were recorded during anesthesia. Renal function was assessed by measuring SCr, BUN, and GFR at 1, 21, and 42 days after surgery. RESULTS: There was no significant difference between groups in measured physiologic variables. No significant difference in GFR, BUN, or SCr between groups or between operated or control kidneys was detected. CONCLUSIONS: Bisection nephrotomy in normal dogs with renal arterial occlusion of 15 minutes and using a simple continuous capsular closure does not adversely affect renal function. CLINICAL RELEVANCE: Bisection nephrotomy, as described in this study, does not decrease renal function; perioperative administration of renoprotective agents is not necessary in normal dogs.     

Molecular differentiation of Hypoderma bovis and Hypoderma lineatum (Diptera,Oestridae) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)

The most variable region of the cytochrome oxidase I (COI) gene of Hypoderma bovis(1) and Hypoderma lineatum(2) (Diptera, Oestridae) was amplified by PCR and the amplicons were sequenced and analysed. PCR products were digested with three restriction enzymes, namely BfaI, HinfI and TaqI, providing informative profiles. H. bovis and H. lineatum sequences revealed an inter-specific variation rate of 8.5%, and an intra-specific variation rate of 0.87 and 0.29%, respectively. The results showed that the COI gene region examined was useful for the differentiation of H. bovis and H. lineatum and that a PCR-RFLP assay is a practical tool for their identification, offering additional diagnostic and epidemiological instruments for the study of cattle grub infestation.     

Efficacy of injectable and pour-on microdose ivermectin in the treatment of goat warble fly infestation by Przhevalskiana silenus (Diptera, Oestridae)

The prophylactic efficacy of microdoses of injectable and pour-on ivermectin formulations against larval stages of Przhevalskiana silenus was assessed in naturally infected goats in the region of Calabria (southern Italy).Sixty-eight goats from two goat farms were divided into five groups: one group remained untreated, while the other four groups were treated with microdoses of ivermectin (5 and 10 microg/kg injectable formulation and 10 and 20 microg/kg pour-on formulation).The microdoses of ivermectin were fully effective in the treatment of goat warble fly infestation (GWFI) as no larvae emerged from the warbles in the treated groups, while all the larvae emerged in the control groups. Irrespective of the type of formulation used, the difference between the treated groups and the control group was statistically significant (P< 0.001). By contrast, no statistical differences were found between the goats treated with the injectable formulation and those receiving the pour-on applications, and between the two doses of the injectable and pour-on formulations used. Given the plasma concentrations it attains at its lowest dose (0.052 - 0.042 ng/ml for the injectable formulation and 0.030 ng/ml for the pour-on) the injectable formulation seems to offer the most reliable route for the administration of ivermectin microdoses and it is acceptable for milk consumption. The introduction of ivermectin in the early eighties and the use of microdoses in some cases have made it possible to control cattle hypodermosis in large areas of Europe. As with cattle hypodermosis, the administration of ivermectin microdoses in goats is particularly interesting because of the low costs involved and the low levels of residues found in goat milk; it may thus constitute the basis for GWFI control campaigns in areas where the disease is prevalent.     

Hemangiosarcoma of the third eyelid in a cat

A case of hemangiosarcoma of the third eyelid in a 15-year-old Domestic Short-haired cat is reported. A small red mass protruding from the anterior surface of the third eyelid in the left eye was present. Only mild bilateral conjunctivitis and nuclear sclerosis were revealed on initial ophthalmic examination. After anti-inflammatory treatment, surgery and cryotherapy were performed. On histopathologic examination the mass was located just beneath the conjunctival epithelium, suggesting that the conjunctiva was the tissue of origin. The tumor showed high cellularity and was composed of pleomorphic spindle and polygonal cells arranged in interlacing bundles and solid sheets. The presence of slit-like spaces between tumor cells containing erythrocytes demonstrated the vascular differentiation. Mitotic activity was moderate, but atypical mitoses were detected. The surgical area healed uneventfully and after 7 months there was no recurrence. To the authors' knowledge, this is the first report of conjunctival hemangiosarcoma of the third eyelid in a cat.